Breast cancer is the second most common cancer among women. Roughly 180,000 women in the U.S. will be diagnosed with malignant breast cancer in 2007-some 40,000 will die. The majority of breast cancers originate in the lobular or ductal cells of the milk-producing glands. In these structures, there are two main cell types: the inner luminal cells surrounded by basal myoepithelium. These cells types are distinct and are the precursors to various forms of breast cancer. As such, it is it is important to study these cell populations independently as well as in co-culture models systems. Current methods for culturing human mammary epithelial cells predominantly select for those of a basal phenotype. Thus, there are no commercial sources of quality-controlled, matched basal and luminal cells (from individual donors) in the U.S., forcing researchers to isolate and characterize the cells on their own. This process is time-consuming, requires access to human tissue, and introduces variation in the preparation and characterization of the cells. Primary cultured human mammary-derived cells are an ideal model currently used to investigate the genesis and understanding of human breast cancer. Because a consistent commercial source of primary human cultured breast-derived luminal and basal cells is unavailable, Zen-Bio, Inc. will address this need by providing a well characterized system to the research community. Phase I of this project focuses on generating an optimized primary human mammary-derived basal/luminal cell system. The Specific Aim is to optimize the isolation and propagation of human mammary epithelial cells followed by characterization of the cellular phenotypes. Optimization of the isolation and propagation will be achieved by systematically modifying aspects of existing protocols to enhance viable cell yields while minimizing population doublings. A detailed analysis of cell properties, including longevity in culture, cell specific gene expression, and cytokeratin production will be performed to initially validate the cell system. These analyses will form the basis of future quality control procedures. Phase II of this project contains two Specific Aims that will generate additional contract services at Zen-Bio. Aim I is to analyze the cytokine and gene expression differences between the two cell types Completion of this Aim leads directly to Aim II which is to establish contract research services at Zen-Bio for those in both the academic as well as pharmaceutical arenas . These services include analysis of cytokine secretion, gene expression, tumor cell(s)-adipose interaction, and cytotoxicity. Multiple product offerings are expected from this proposal: a matched, primary human mammary cell system, support media and reagents, contract assay services, and kits for basal/luminal cell research. PUBLIC HEALTH RELEVANCE: At the completion of this project, a commercially available, fully characterized human, primary mammary- derived basal/luminal cell system and support reagents will be offered to researchers. The accessibility of this currently unavailable system will not only provide a greater opportunity to investigate novel methods to further understand the etiology of breast cancer but also to investigate potential cures. [unreadable] [unreadable] [unreadable]